Croatian veteran Karlovic – who celebrated his 40th birthday on February 28 – edged Matthew Ebden in straight sets on Thursday.
Karlovic became the first 40-year-old since Jimmy Connors in Halle 24 years ago to win on the Tour.
Indian Prajnesh Gunneswaran, meanwhile, enjoyed a memorable ATP Masters 1000 debut against Benoit Paire.
Karlovic defying Father Time
Making his 14th Indian Wells appearance, Karlovic outlasted Ebden 7-6 (7-3) 7-6 (7-3).
Karlovic – who became the oldest ATP Tour finalist since 1977 in Pune this year – fired down 18 aces in a match lasting one hour, 49 minutes.
A 2011 Indian Wells quarter-finalist, Karlovic will meet countryman Borna Coric in the second round.
Gunneswaran stuns Paire
The 29-year-old upstaged former world number 18 Paire 7-6 (7-5) 6-4 for just his second main-draw victory.
Making his first appearance in a Masters main-draw match, Gunneswaran was too good for the Frenchman as he booked a second-round clash with Nikoloz Basilashvili.
He saved five of eight break points against Paire, who had a first-serve percentage of just 38.
Giron shocks Chardy
Another qualifier Marcos Giron produced one of the biggest upsets of the day.
Giron stunned Jeremy Chardy 7-6 (7-3) 7-6 (7-1) in his Masters debut on Thursday.
Jordan Thompson will meet Dominic Thiem in the second round after surviving a three-hour marathon against Federico Delbonis 6-4 6-7 (6-8) 7-6 (7-5) for his maiden Masters triumph, while Leonardo Mayer beat Reilly Opelka in straight sets.
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